bims-tuchim 2021-10-17 papers

bims-tuchim

Biomed News

on Tumor-on-chip models

Issue of 2021–10–17
fourteen papers selected by
Philipp Albrecht, Friedrich Schiller University

Novel 3D µtissues Mimicking the Fibrotic Stroma in Pancreatic Cancer to Study Cellular Interactions and Stroma-Modulating Therapeutics.
Using Organotypic Tissue Slices to Investigate the Microenvironment of Pancreatic Cancer: Pharmacotyping and Beyond.
Microfluidic Arrays of Breast Tumor Spheroids for Drug Screening and Personalized Cancer Therapies.
Pancreatic Cancer Microenvironment and Cellular Composition: Current Understandings and Therapeutic Approaches.
Angiogenic Sprouting Dynamics Mediated by Endothelial-Fibroblast Interactions in Microfluidic Systems.
The Heterogeneity of the Tumor Microenvironment as Essential Determinant of Development, Progression and Therapy Response of Pancreatic Cancer.
The Cellular Origins of Cancer-Associated Fibroblasts and Their Opposing Contributions to Pancreatic Cancer Growth.
High-Throughput 3D In Vitro Tumor Vasculature Model for Real-Time Monitoring of Immune Cell Infiltration and Cytotoxicity.
Understanding and Improving Cellular Immunotherapies Against Cancer: From Cell-Manufacturing to Tumor-Immune Models.
The Diverse Applications of Pancreatic Ductal Adenocarcinoma Organoids.
Spatially confined sub-tumor microenvironments in pancreatic cancer.
Heterogeneity in Pancreatic Cancer Fibroblasts-TGFβ as a Master Regulator?
Peristaltic on-chip pump for tunable media circulation and whole blood perfusion in PDMS-free organ-on-chip and Organ-Disc systems.
Adoption of organ-on-chip platforms by the pharmaceutical industry.

Cancers (Basel). 2021 Oct 06. pii: 5006. [Epub ahead of print]13(19):

Novel 3D µtissues Mimicking the Fibrotic Stroma in Pancreatic Cancer to Study Cellular Interactions and Stroma-Modulating Therapeutics.

Kunal P Pednekar, Marcel A Heinrich, Joop van Baarlen, Jai Prakash.

  Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive tumor type with low patient survival due to the low efficacy of current treatment options. Cancer-associated fibroblasts (CAFs) in the tumor microenvironment (TME) create a dense fibrotic environment around the tumor cells, preventing therapies from reaching their target. Novel 3D in vitro models are needed that mimic this fibrotic barrier for the development of therapies in a biologically relevant environment. Here, novel PDAC microtissues (µtissues) consisting of pancreatic cancer cell core surrounded by a CAF-laden collagen gel are presented, that is based on the cells own contractility to form a hard-to-penetrate barrier. The contraction of CAFs is demonstrated facilitating the embedding of tumor cells in the center of the µtissue as observed in patients. The µtissues displayed a PDAC-relevant gene expression by comparing their gene profile with transcriptomic patient data. Furthermore, the CAF-dependent proliferation of cancer cells is presented, as well as the suitability of the µtissues to serve as a platform for the screening of CAF-modulating therapies in combination with other (nano)therapies. It is envisioned that these PDAC µtissues can serve as a high-throughput platform for studying cellular interactions in PDAC and for evaluating different treatment strategies in the future.

Keywords:  3D in-vitro model; 3D µtissues; cancer-associated fibroblasts; cell contraction; collagen hydrogel; pancreatic ductal adenocarcinoma; primary pancreatic stellate cells; tumor microenvironment

DOI:  https://doi.org/10.3390/cancers13195006
Cancers (Basel). 2021 Oct 05. pii: 4991. [Epub ahead of print]13(19):

Using Organotypic Tissue Slices to Investigate the Microenvironment of Pancreatic Cancer: Pharmacotyping and Beyond.

Jonathan Robert Weitz, Herve Tiriac, Tatiana Hurtado de Mendoza, Alexis Wascher, Andrew M Lowy.

  Organotypic tissue slices prepared from patient tumors are a semi-intact ex vivo preparation that recapitulates many aspects of the tumor microenvironment (TME). While connections to the vasculature and nervous system are severed, the integral functional elements of the tumor remain intact for many days during the slice culture. During this window of time, the slice platforms offer a suite of molecular, biomechanical and functional tools to investigate PDAC biology. In this review, we first briefly discuss the development of pancreatic tissue slices as a model system. Next, we touch upon using slices as an orthogonal approach to study the TME as compared to other established 3D models, such as organoids. Distinct from most other models, the pancreatic slices contain autologous immune and other stromal cells. Taking advantage of the existing immune cells within the slices, we will discuss the breakthrough studies which investigate the immune compartment in the pancreas slices. These studies will provide an important framework for future investigations seeking to exploit or reprogram the TME for cancer therapy.

Keywords:  3D culture; PDAC; microenvironment; organotypic; pancreatic cancer; slices

DOI:  https://doi.org/10.3390/cancers13194991
Adv Healthc Mater. 2021 Oct 11. e2101085

Microfluidic Arrays of Breast Tumor Spheroids for Drug Screening and Personalized Cancer Therapies.

Elisabeth Prince, Sina Kheiri, Yihe Wang, Fei Xu, Jennifer Cruikshank, Valentina Topolskaia, Huachen Tao, Edmund W K Young, Alison P McGuigan, David Cescon, Eugenia Kumacheva.

  One of the obstacles limiting progress in the development of effective cancer therapies is the shortage of preclinical models that capture the dynamic nature of tumor microenvironments. Interstitial flow strongly impacts tumor response to chemotherapy; however, conventional in vitro cancer models largely disregard this key feature. Here, we report a proof of principle microfluidic platform for the generation of large arrays of breast tumor spheroids that are grown under close-to-physiological flow in a biomimetic hydrogel. We use this cancer spheroids-on-a-chip model for time- and labor-efficient studies of the effects of drug dose and supply rate on the chemosensitivity of breast tumor spheroids. We show the capability to grow large arrays of tumor spheroids from patient-derived cells of different breast cancer subtypes and demonstrate correlation between in vivo drug efficacy and on-chip spheroid drug response. The proposed platform can serve as an in vitro preclinical model for the development of personalized cancer therapies and effective screening of new anticancer drugs. This article is protected by copyright. All rights reserved.

Keywords:  Breast cancer; cancer; drug testing; microfluidics; personalized medicine; preclinical models; spheroids

DOI:  https://doi.org/10.1002/adhm.202101085
Cancers (Basel). 2021 Oct 08. pii: 5028. [Epub ahead of print]13(19):

Pancreatic Cancer Microenvironment and Cellular Composition: Current Understandings and Therapeutic Approaches.

Linh-Huyen Truong, Siim Pauklin.

  Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal human solid tumors, despite great efforts in improving therapeutics over the past few decades. In PDAC, the distinct characteristic of the tumor microenvironment (TME) is the main barrier for developing effective treatments. PDAC TME is characterized by a dense stroma, cancer-associated fibroblasts, and immune cells populations that crosstalk to the subpopulations of neoplastic cells that include cancer stem cells (CSCs). The heterogeneity in TME is also exhibited in the diversity and dynamics of acellular components, including the Extracellular matrix (ECM), cytokines, growth factors, and secreted ligands to signaling pathways. These contribute to drug resistance, metastasis, and relapse in PDAC. However, clinical trials targeting TME components have often reported unexpected results and still have not benefited patients. The failures in those trials and various efforts to understand the PDAC biology demonstrate the highly heterogeneous and multi-faceted TME compositions and the complexity of their interplay within TME. Hence, further functional and mechanistic insight is needed. In this review, we will present a current understanding of PDAC biology with a focus on the heterogeneity in TME and crosstalk among its components. We also discuss clinical challenges and the arising therapeutic opportunities in PDAC research.

Keywords:  cancer therapy; fibroblasts; immune cells; pancreatic ductal adenocarcinoma; tumor microenvironment; tumor stroma

DOI:  https://doi.org/10.3390/cancers13195028
Adv Biol (Weinh). 2021 Oct 15. e2101080

Angiogenic Sprouting Dynamics Mediated by Endothelial-Fibroblast Interactions in Microfluidic Systems.

Noosheen Walji, Sina Kheiri, Edmond W K Young.

  Angiogenesis, the development of new blood vessels from existing vasculature, is a key process in normal development and pathophysiology. In vitro models are necessary for investigating mechanisms of angiogenesis and developing antiangiogenic therapies. Microfluidic cell culture models of angiogenesis are favored for their ability to recapitulate 3D tissue structures and control spatiotemporal aspects of the microenvironments. To capture the angiogenesis process, microfluidic models often include endothelial cells and a fibroblast component. However, the influence of fibroblast organization on resulting angiogenic behavior remains unclear. Here a comparative study of angiogenic sprouting on a microfluidic chip induced by fibroblasts in 2D monolayer, 3D dispersed, and 3D spheroid culture formats, is conducted. Vessel morphology and sprout distribution for each configuration are measured, and these observations are correlated with measurements of secreted factors and numerical simulations of diffusion gradients. The results demonstrate that angiogenic sprouting varies in response to fibroblast organization with correlating variations in secretory profile and secreted factor gradients across the microfluidic device. This study is anticipated to shed light on how sprouting dynamics are mediated by fibroblast configuration such that the microfluidic cell culture design process includes the selection of a fibroblast component where the effects are known and leveraged.

Keywords:  angiogenesis; angiogenic sprouting; fibroblasts; microfluidics; organ-on-a-chip; tumor microenvironment

DOI:  https://doi.org/10.1002/adbi.202101080
Cancers (Basel). 2021 Sep 30. pii: 4932. [Epub ahead of print]13(19):

The Heterogeneity of the Tumor Microenvironment as Essential Determinant of Development, Progression and Therapy Response of Pancreatic Cancer.

Group Young Researchers In Inflammatory Carcinogenesis, Anna Maxi Wandmacher, Anne-Sophie Mehdorn, Susanne Sebens.

  Pancreatic ductal adenocarcinoma (PDAC) is commonly diagnosed at advanced stages and most anti-cancer therapies have failed to substantially improve prognosis of PDAC patients. As a result, PDAC is still one of the deadliest tumors. Tumor heterogeneity, manifesting at multiple levels, provides a conclusive explanation for divergent survival times and therapy responses of PDAC patients. Besides tumor cell heterogeneity, PDAC is characterized by a pronounced inflammatory stroma comprising various non-neoplastic cells such as myofibroblasts, endothelial cells and different leukocyte populations which enrich in the tumor microenvironment (TME) during pancreatic tumorigenesis. Thus, the stromal compartment also displays a high temporal and spatial heterogeneity accounting for diverse effects on the development, progression and therapy responses of PDAC. Adding to this heterogeneity and the impact of the TME, the microbiome of PDAC patients is considerably altered. Understanding this multi-level heterogeneity and considering it for the development of novel therapeutic concepts might finally improve the dismal situation of PDAC patients. Here, we outline the current knowledge on PDAC cell heterogeneity focusing on different stromal cell populations and outline their impact on PDAC progression and therapy resistance. Based on this information, we propose some novel concepts for treatment of PDAC patients.

Keywords:  CAFs; PDAC; endothelial cells; immune cells; macrophages; microbiome; myofibroblasts; personalized medicine; tumor stroma

DOI:  https://doi.org/10.3390/cancers13194932
Front Cell Dev Biol. 2021 ;9 743907

The Cellular Origins of Cancer-Associated Fibroblasts and Their Opposing Contributions to Pancreatic Cancer Growth.

Paul Manoukian, Maarten Bijlsma, Hanneke van Laarhoven.

  Pancreatic tumors are known to harbor an abundant and highly desmoplastic stroma. Among the various cell types that reside within tumor stroma, cancer-associated fibroblasts (CAFs) have gained a lot of attention in the cancer field due to their contributions to carcinogenesis and tumor architecture. These cells are not a homogeneous population, but have been shown to have different origins, phenotypes, and contributions. In pancreatic tumors, CAFs generally emerge through the activation and/or recruitment of various cell types, most notably resident fibroblasts, pancreatic stellate cells (PSCs), and tumor-infiltrating mesenchymal stem cells (MSCs). In recent years, single cell transcriptomic studies allowed the identification of distinct CAF populations in pancreatic tumors. Nonetheless, the exact sources and functions of those different CAF phenotypes remain to be fully understood. Considering the importance of stromal cells in pancreatic cancer, many novel approaches have aimed at targeting the stroma but current stroma-targeting therapies have yielded subpar results, which may be attributed to heterogeneity in the fibroblast population. Thus, fully understanding the roles of different subsets of CAFs within the stroma, and the cellular dynamics at play that contribute to heterogeneity in CAF subsets may be essential for the design of novel therapies and improving clinical outcomes. Fortunately, recent advances in technologies such as microfluidics and bio-printing have made it possible to establish more advanced ex vivo models that will likely prove useful. In this review, we will present the different roles of stromal cells in pancreatic cancer, focusing on CAF origin as a source of heterogeneity, and the role this may play in therapy failure. We will discuss preclinical models that could be of benefit to the field and that may contribute to further clinical development.

Keywords:  cancer-associated fibroblasts; cellular origins; heterogeneity; pancreatic ductal adenocarcinoma; pre-clinical models; radiation; resistance; stroma

DOI:  https://doi.org/10.3389/fcell.2021.743907
Front Immunol. 2021 ;12 733317

High-Throughput 3D In Vitro Tumor Vasculature Model for Real-Time Monitoring of Immune Cell Infiltration and Cytotoxicity.

Jiyoung Song, Hyeri Choi, Seung Kwon Koh, Dohyun Park, James Yu, Habin Kang, Youngtaek Kim, Duck Cho, Noo Li Jeon.

  Recent advances in anticancer therapy have shown dramatic improvements in clinical outcomes, and adoptive cell therapy has emerged as a type of immunotherapy that can modulate immune responses by transferring engineered immune cells. However, a small percentage of responders and their toxicity remain as challenges. Three-dimensional (3D) in vitro models of the tumor microenvironment (TME) have the potential to provide a platform for assessing and predicting responses to therapy. This paper describes an in vitro 3D tumor model that incorporates clusters of colorectal cancer (CRC) cells around perfusable vascular networks to validate immune-cell-mediated cytotoxicity against cancer cells. The platform is based on an injection-molded 3D co-culture model and composed of 28 microwells where separate identical vascularized cancer models can be formed. It allows robust hydrogel patterning for 3D culture that enables high-throughput experimentation. The uniformity of the devices resulted in reproducible experiments that allowed 10× more experiments to be performed when compared to conventional polydimethylsiloxane (PDMS)-based microfluidic devices. To demonstrate its capability, primary natural killer (NK) cells were introduced into the vascularized tumor network, and their activities were monitored using live-cell imaging. Extravasation, migration, and cytotoxic activity against six types of CRC cell lines were tested and compared. The consensus molecular subtypes (CMS) of CRC with distinct immune responses resulted in the highest NK cell cytotoxicity against CMS1 cancer cells. These results show the potential of our vascularized tumor model for understanding various steps involved in the immune response for the assessment of adoptive cell therapy.

Keywords:  NK cell cytotoxicity; adoptive cell therapy; live-cell imaging; microfluidics; tumor vasculature

DOI:  https://doi.org/10.3389/fimmu.2021.733317
Adv Drug Deliv Rev. 2021 Oct 12. pii: S0169-409X(21)00396-3. [Epub ahead of print] 114003

Understanding and Improving Cellular Immunotherapies Against Cancer: From Cell-Manufacturing to Tumor-Immune Models.

Rachel Ringquist, Delta Ghoshal, Ritika Jain, Krishnendu Roy.

  The tumor microenvironment (TME) is shaped by dynamic metabolic and immune interactions between precancerous and cancerous tumor cells and stromal cells like epithelial cells, fibroblasts, endothelial cells, and hematopoietically-derived immune cells. The metabolic states of the TME, including the hypoxic and acidic niches, influence the immunosuppressive phenotypes of the stromal and immune cells, which confers resistance to both host-mediated tumor killing and therapeutics. Numerous in vitro TME platforms for studying immunotherapies, including cell therapies, are being developed. However, we do not yet understand which immune and stromal components are most critical and how much model complexity is needed to answer specific questions. In addition, scalable sourcing and quality-control of appropriate TME cells for reproducibly manufacturing these platforms remain challenging. In this regard, lessons from the manufacturing of immunomodulatory cell therapies could provide helpful guidance. Although immune cell therapies have shown unprecedented results in hematological cancers and hold promise in solid tumors, their manufacture poses significant scale, cost, and quality control challenges. This review first provides an overview of the in vivo TME, discussing the most influential cell populations in the tumor-immune landscape. We then evaluate current immune-tumor models of TME and immunotherapies, highlighting the complexity, architecture, function, and cell sources. Next, we summarize current approaches for cell therapies against cancers and the relevant manufacturing platforms. Finally, we present the technical and fundamental knowledge gaps in both cell manufacturing systems and immune-TME models that must be addressed to elucidate the interactions between endogenous tumor immunity and exogenous engineered immunity.

Keywords:  CAR; TME; bioprinting; cancer; immunotherapy; organoid; spheroid; tumor-on-chip

DOI:  https://doi.org/10.1016/j.addr.2021.114003
Cancers (Basel). 2021 Oct 04. pii: 4979. [Epub ahead of print]13(19):

The Diverse Applications of Pancreatic Ductal Adenocarcinoma Organoids.

Ronnie Ren Jie Low, Wei Wen Lim, Paul M Nguyen, Belinda Lee, Michael Christie, Antony W Burgess, Peter Gibbs, Sean M Grimmond, Frédéric Hollande, Tracy L Putoczki.

  Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal solid malignancies. While immortalized cancer cell lines and genetically engineered murine models have increased our understanding of PDAC tumorigenesis, they do not recapitulate inter- and intra-patient heterogeneity. PDAC patient derived organoid (PDO) biobanks have overcome this hurdle, and provide an opportunity for the high throughput screening of potential new therapies. This review provides a summary of the PDAC PDO biobanks established to date, and discusses how they have advanced our understanding of PDAC biology. Looking forward, the development of coculturing techniques for specific immune or stromal cell populations will enable a better understanding of the crosstalk that occurs within the tumor microenvironment, and the impact of this crosstalk on treatment response.

Keywords:  PDAC; cancer; coculture; drug screen; fibroblast; immune; microenvironment; organoid; patient derived; therapeutic

DOI:  https://doi.org/10.3390/cancers13194979
Cell. 2021 Oct 07. pii: S0092-8674(21)01105-3. [Epub ahead of print]

Spatially confined sub-tumor microenvironments in pancreatic cancer.

Barbara T Grünwald, Antoine Devisme, Geoffroy Andrieux, Foram Vyas, Kazeera Aliar, Curtis W McCloskey, Andrew Macklin, Gun Ho Jang, Robert Denroche, Joan Miguel Romero, Prashant Bavi, Peter Bronsert, Faiyaz Notta, Grainne O'Kane, Julie Wilson, Jennifer Knox, Laura Tamblyn, Molly Udaskin, Nikolina Radulovich, Sandra E Fischer, Melanie Boerries, Steven Gallinger, Thomas Kislinger, Rama Khokha.

  Intratumoral heterogeneity is a critical frontier in understanding how the tumor microenvironment (TME) propels malignant progression. Here, we deconvolute the human pancreatic TME through large-scale integration of histology-guided regional multiOMICs with clinical data and patient-derived preclinical models. We discover "subTMEs," histologically definable tissue states anchored in fibroblast plasticity, with regional relationships to tumor immunity, subtypes, differentiation, and treatment response. "Reactive" subTMEs rich in complex but functionally coordinated fibroblast communities were immune hot and inhabited by aggressive tumor cell phenotypes. The matrix-rich "deserted" subTMEs harbored fewer activated fibroblasts and tumor-suppressive features yet were markedly chemoprotective and enriched upon chemotherapy. SubTMEs originated in fibroblast differentiation trajectories, and transitory states were notable both in single-cell transcriptomics and in situ. The intratumoral co-occurrence of subTMEs produced patient-specific phenotypic and computationally predictable heterogeneity tightly linked to malignant biology. Therefore, heterogeneity within the plentiful, notorious pancreatic TME is not random but marks fundamental tissue organizational units.

Keywords:  cancer-associated fibroblasts; pancreatic cancer; patient-derived organoids; proteomics; stromal heterogeneity; systems biology; treatment resistance; tumor microenvironment

DOI:  https://doi.org/10.1016/j.cell.2021.09.022
Cancers (Basel). 2021 Oct 04. pii: 4984. [Epub ahead of print]13(19):

Heterogeneity in Pancreatic Cancer Fibroblasts-TGFβ as a Master Regulator?

Dale M Watt, Jennifer P Morton.

  Pancreatic ductal adenocarcinoma is an aggressive disease for which there are very few available therapies. It is notable for its high degree of tumour complexity, with the tumour microenvironment often accounting for the majority of the tumour volume. Until recently, the biology of the stroma was poorly understood, particularly in terms of heterogeneity. Recent research, however, has shed light on the intricacy of signalling within the stroma and particularly the molecular and functional heterogeneity of the cancer associated fibroblasts. In this review, we summarise the recent improvements in our understanding of the different fibroblast populations within PDAC, with a focus on the role TGFβ plays to dictate their formation and function. These studies have highlighted some of the reasons for the failure of trials targeting the tumour stroma, however, there are still considerable gaps in our knowledge, and more work is needed to make effective fibroblast targeting a reality in the clinic.

Keywords:  CAFs; TGFβ; fibroblasts; pancreatic cancer; stroma

DOI:  https://doi.org/10.3390/cancers13194984
Lab Chip. 2021 Oct 12. 21(20): 3963-3978

Peristaltic on-chip pump for tunable media circulation and whole blood perfusion in PDMS-free organ-on-chip and Organ-Disc systems.

Stefan Schneider, Marvin Bubeck, Julia Rogal, Huub J Weener, Cristhian Rojas, Martin Weiss, Michael Heymann, Andries D van der Meer, Peter Loskill.

Organ-on-chip (OoC) systems have become a promising tool for personalized medicine and drug development with advantages over conventional animal models and cell assays. However, the utility of OoCs in industrial settings is still limited, as external pumps and tubing for on-chip fluid transport are dependent on error-prone, manual handling. Here, we present an on-chip pump for OoC and Organ-Disc systems, to perfuse media without external pumps or tubing. Peristaltic pumping is implemented through periodic compression of a flexible pump layer. The disc-shaped, microfluidic module contains four independent systems, each lined with endothelial cells cultured under defined, peristaltic perfusion. Both cell viability and functionality were maintained over several days shown by supernatant analysis and immunostaining. Integrated, on-disc perfusion was further used for cytokine-induced cell activation with physiologic cell responses and for whole blood perfusion assays, both demonstrating the versatility of our system for OoC applications.

DOI: https://doi.org/10.1039/d1lc00494h
Nat Rev Drug Discov. 2021 Oct 13.

Adoption of organ-on-chip platforms by the pharmaceutical industry.

Paul Vulto, Jos Joore.

DOI: https://doi.org/10.1038/s41573-021-00323-0